Anti-CBLB antibody [246C5a] (ab54362)
![Anti-CBLB antibody [246C5a] (ab54362)](https://www.abcam.com/ps/products/54/ab54362/Images/ab54362-67202-anti-cblb-antibody-246c5a-immunohistochemistry.jpg "Anti-CBLB antibody [246C5a] (ab54362)")
Key features and details
- Mouse monoclonal [246C5a] to CBLB
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG2a
Overview
-
Product name
Anti-CBLB antibody [246C5a]
See all CBLB primary antibodies -
Description
Mouse monoclonal [246C5a] to CBLB -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB HumanRecombinant fragment
-
Immunogen
Recombinant C terminal fragment of Human CBLB.
-
Positive control
- WB: CTLL-2, HeLa and MCF7 cell lysates
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. -
Storage buffer
pH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 1% BSA, PBS -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
ab54362 was purified using protein G column chromatography from culture supernatant of hybridoma cultured in a medium containing bovine IgG depleted (approximately 95%) fetal bovine serum and filtered through a 0.22µm membrane. -
Clonality
Monoclonal -
Clone number
246C5a -
Isotype
IgG2a -
Research areas
Images
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBLB antibody [246C5a] (ab54362)
IHC image of ab54362 staining in human T-cell lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab54362, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
![Western blot - Anti-CBLB antibody [246C5a] (ab54362)](https://www.abcam.com/ps/products/54/ab54362/Images/ab54362-383742-anti-cblb-antibody-246c5a-western-blot-hela-ctll2-mcf7.jpg)
Western blot - Anti-CBLB antibody [246C5a] (ab54362)All lanes : Anti-CBLB antibody [246C5a] (ab54362) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CBLB knockout HeLa cell lysate
Lane 3 : CTLL-2 cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 109 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab54362 observed at 130 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab54362 Anti-CBLB antibody [246C5a] was shown to specifically react with CBLB in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264711 (knockout cell lysate ab258342) was used. Wild-type and CBLB knockout samples were subjected to SDS-PAGE. ab54362 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
-
![Western blot - Anti-CBLB antibody [246C5a] (ab54362)](https://www.abcam.com/ps/products/54/ab54362/Images/ab54362-67203-anti-cblb-antibody-246c5a-western-blot.gif)
Western blot - Anti-CBLB antibody [246C5a] (ab54362)Anti-CBLB antibody [246C5a] (ab54362) + immunizing recombinant protein
Predicted band size: 109 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
The molecular weight of the band on the Western blot does not correspond to the molecular weight of the natural protein as the immunizing recombinant protein fragment was used as the test antigen.
